519 .
SYNTROPHOMONAS MEDIUM (for Syntrophospora cellicola 19J-3)
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Equipment needed:
Autoclave
Filter
Final pH:
7.2
Select a strain (optional):
Solution A #
902
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Mineral solution | 50.0 | ml | ||
|
|
FeCl2 solution | 1.0 | ml | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
Rumen fluid
, clarified
(see Medium No. 266) |
50.0 | ml | ||
|
|
Trypticase peptone
(BD-BBL) |
1.0 | g | ||
|
|
Vitamin solution | 5.0 | ml | ||
|
|
Butyric acid | 1.7 | g | ||
|
|
Na2SO4 | 2.8 | g | ||
|
|
Resazurin | 1.0 | mg | ||
|
|
Distilled water | 795.0 | ml | ||
| 1 Mix components, adjust pH to 7.2, bring to a boil for a few seconds and cool down under a stream of a N2-CO2 (80:20) gas mixture. Distribute Solution A into culture vessels (e.g., 9 ml per tube) under the same gas mixture, seal with butyl rubber stoppers and autoclave. | |||||
| 2 To complete the medium, add 0.1 volume of 3.5% NaHCO3 solution [filter-sterilized and stored under a N2-CO2 (80:20) gas mixture] and reduce the medium with 0.3 g/L (final) L-cysteine x HCl x H2O and 0.3 g/L (final) Na2S x 9H2O (autoclave and store 3% stock solutions under a N2 atmosphere). | |||||
Mineral solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 10.0 | g |
|
|
MgCl2 x 6 H2O | 6.6 | g |
|
|
NaCl | 8.0 | g |
|
|
NH4Cl | 8.0 | g |
|
|
CaCl2 x 2 H2O | 1.0 | g |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 0.25 | mg |
|
|
Nicotinic acid | 2.50 | mg |
|
|
Thiamine HCl | 1.25 | mg |
|
|
p-Aminobenzoic acid | 1.25 | mg |
|
|
DL-Calcium pantothenate | 0.62 | mg |
|
|
Pyridoxine hydrochloride | 6.20 | mg |
|
|
Distilled water | 1000.00 | ml |
FeCl2 solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
|
FeCl2 x 4 H2O | 1.5 | g |
|
|
Distilled water | 990.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnCl2 | 70 | mg |
|
|
MnCl2 x 4 H2O | 100 | mg |
|
|
H3BO3 | 6 | mg |
|
|
CoCl2 x 6 H2O | 190 | mg |
|
|
CuCl2 x 2 H2O | 2 | mg |
|
|
NiCl2 x 6 H2O | 24 | mg |
|
|
Na2MoO4 x 2 H2O | 36 | mg |
|
|
Distilled water | 1000 | ml |